CRF-like effects of sauvagine and urotensin I on synaptosomal tyrosine hydroxylase activity of mouse striatum.

In the present study we have investigated whether sauvagine (SVG) and urotensin I (UT), two peptides displaying sequence homology with corticotropin-releasing factor (CRF), could affect synaptosomal tyrosine hydroxylase (TH) activity of mouse striatum in a manner similar to CRF. The enzyme activity was assayed in supernatants obtained following sonication and centrifugation of homogenates preincubated with the peptides. SVG and UT produced a concentration-dependent increase of TH activity with a half-maximal effect obtained at 5 and 10 nM, respectively. SVG and UT were as effective as CRF with maximal stimulations corresponding to 52-58% increase of basal enzyme activity, whereas the rank order of potency was SVG greater than UT = CRF. Kinetic analysis of TH activity versus low concentrations of the pterin co-factor (0.05-0.4 mM) indicated that the stimulations elicited by CRF, SVG and UT were associated with an increase in the Vmax of the enzyme form with high affinity for the co-factor. The CRF receptor antagonist alpha-helical CRF9-41 inhibited the effects of all 3 peptides. Moreover, the combined addition of CRF with either SVG or UT did not produce additive effects on TH activity. The stimulatory effects of CRF, SVG and UT were dependent on the concentration of extracellular free Ca2+, being minimal in a Ca2(+)-free medium and maximal at about 0.5 mM extracellular free Ca2+. These results indicate that SVG and UT can mimic the effect of CRF on synaptosomal TH by acting on a common receptor site associated with a Ca2(+)-dependent mechanism.